Abstract
BackgroundThe glassy-winged sharpshooter Homalodisca vitripennis (Hemiptera: Cicadellidae), is a xylem-feeding leafhopper and important vector of the bacterium Xylella fastidiosa; the causal agent of Pierce’s disease of grapevines. The functional complexity of the transcriptome of H. vitripennis has not been elucidated thus far. It is a necessary blueprint for an understanding of the development of H. vitripennis and for designing efficient biorational control strategies including those based on RNA interference.ResultsHere we elucidate and explore the transcriptome of adult H. vitripennis using high-throughput paired end deep sequencing and de novo assembly. A total of 32,803,656 paired-end reads were obtained with an average transcript length of 624 nucleotides. We assembled 32.9 Mb of the transcriptome of H. vitripennis that spanned across 47,265 loci and 52,708 transcripts. Comparison of our non-redundant database showed that 45% of the deduced proteins of H. vitripennis exhibit identity (e-value ≤1−5) with known proteins. We assigned Gene Ontology (GO) terms, Kyoto Encyclopedia of Genes and Genomes (KEGG) annotations, and potential Pfam domains to each transcript isoform. In order to gain insight into the molecular basis of key regulatory genes of H. vitripennis, we characterized predicted proteins involved in the metabolism of juvenile hormone, and biogenesis of small RNAs (Dicer and Piwi sequences) from the transcriptomic sequences. Analysis of transposable element sequences of H. vitripennis indicated that the genome is less expanded in comparison to many other insects with approximately 1% of the transcriptome carrying transposable elements.ConclusionsOur data significantly enhance the molecular resources available for future study and control of this economically important hemipteran. This transcriptional information not only provides a more nuanced understanding of the underlying biological and physiological mechanisms that govern H. vitripennis, but may also lead to the identification of novel targets for biorationally designed control strategies.
Highlights
The transcriptome is a complete set of RNA transcripts produced inside an organism at a particular time, and represents a genomic blueprint of that organism
Unlocking the complexity of the transcriptome is essential for interpreting the functional elements of a genome which can be applied for more effective downstream applications including developing genome based biorational control strategies such as those based on RNA interference
Maintenance of H. vitripennis and RNA Extraction A colony of H. vitripennis was maintained at the University of California-Davis Contained Research Facility (CRF) in cages containing a mixture of host plants as previously described [20]
Summary
The transcriptome is a complete set of RNA transcripts produced inside an organism at a particular time, and represents a genomic blueprint of that organism. H. vitripennis is a very economically important pest of a wide range of plants including Citrus spp., grapes (Vitis vinifera), and almonds (Prunus dulcis) [20] This insect serves as a very robust vector of the bacterium Xylella fastidiosa, the causal agent of Pierce’s disease of grapevines and citrus variegated chlorosis disease [21]. RNAi is a widely used tool to knock down and analyze the function of genes, especially in non-model organisms where the systematic recovery of mutants is not feasible This approach, is limited in H. vitripennis because of the lack of information of potential targets. Robust systemic RNAi is absent in insects [27] because they lack a functional RNA dependent RNA polymerase (RdRP) that amplifies small RNAs. The glassy-winged sharpshooter Homalodisca vitripennis (Hemiptera: Cicadellidae), is a xylem-feeding leafhopper and important vector of the bacterium Xylella fastidiosa; the causal agent of Pierce’s disease of grapevines. It is a necessary blueprint for an understanding of the development of H. vitripennis and for designing efficient biorational control strategies including those based on RNA interference
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